Protease Factor Xa ?

Steve Minchin S.D.Minchin at bham.ac.uk
Wed Jun 29 09:25:18 EST 1994


In article <28JUN199420073552 at opal.tufts.edu> jpalek at opal.tufts.edu (HYNEK WICHTERLE) writes:
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>From: jpalek at opal.tufts.edu (HYNEK WICHTERLE)
>Subject: Protease Factor Xa ?
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>Does anybody know how to use Factor Xa as a protease? (concentration,
>conditions...). According to Biochemistry Labfax, it should cut Arg-X of
>Gly-Arg-X.

>Could you help me please, also with conditions for cutting Asn-Gly with
>Hydroxilamine.

>thank you in advance

>hynek wichterle (jpalek at opal.tufts.edu)

Hi,

We have found that there is no standard conditions for Factor Xa digestion. 
Cleavage will be influenced by the accessability of the cleavage site and the 
surrounding peptide sequence.

You should therefore optimise the conditions for your particular protein.

Things to vary are:

pH (7-10)
Substrate Conc. 0.1-20mg/ml
enzyme:Substrate ratio (w:w) 1:10 - 1:200
temperature 0-37 degrees C
time 1- 20 hours

There are several references including Nagia & Thogersen (1984) Nature Vol.309 
pg810-812.

All the Best

Steve Minchin

 
---------------------------------------------------------
Dr. Steve Minchin,

School of Biochemistry, The University of Birmingham,
Edgbaston, Birmingham, B15 2TT, U.K.

E-Mail
InterNet:       S.D.Minchin at bham.ac.uk
Janet:          S.D.Minchin at uk.ac.bham

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